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目的鉴定原发性胆汁性肝硬化(PBC)患者中自身抗原丙酮酸脱氢酶(PDC-E2)上的HLA-A觹0201限制性CD8+CTL表位,为临床探索特异性免疫治疗奠定基础。方法应用数据库SYFPEITHI预测PDC-E2上两段涵盖B细胞表位和CD4+T细胞表位的氨基酸序列(163～184aa和36～49aa)附近可能存在的HLA-A觹0201限制性T细胞表位,再通过T2细胞株、细胞增殖试验和细胞毒性检测分别分析各抗原肽与HLA-A觹0201的结合力、诱导患者外周血单个核细胞(PBMCs)增殖能力及抗原肽诱导的抗原特异性T细胞杀伤毒性,逐步鉴定HLA-A觹0201限制性CD8+CTL细胞表位。结果数据库初步预测到5个可能性较大的HLA-A觹0201限制性抗原肽,其中两个抗原肽(159～167aa和165～174aa)显示与T2细胞上HLA-A觹0201分子有较高的亲和力,这两个抗原肽能刺激大部分HLA-A觹0201阳性PBC患者PBMC增殖,并且由其诱导产生的CTL具有特异杀伤活性。结论位于PDC-E2内酯酰区上的KLSEGDLLA穴159～167aa)和LLAEIETDKA穴165～174aa雪是PBC患者体内HLA-A觹0201限制性的CD8+CTL表位。
Objective To identify HLA-A 觹 0201-restricted CD8 + CTL epitopes on the autoantigen pyruvate dehydrogenase (PDC-E2) in patients with primary biliary cirrhosis (PBC) and lay the foundation for clinical exploration of specific immunotherapy . Methods The SYFPEITHI database was used to predict the possible HLA-A 觹 0201-restricted T cell epitopes in the two segments of PDC-E2 covering the amino acid sequences of B cell epitopes and CD4 + T cell epitopes (163-184aa and 36-49aa) , And then through the T2 cell line, cell proliferation test and cytotoxicity assay were analyzed for each antigen peptide and HLA-A 觹 0201 binding capacity to induce peripheral blood mononuclear cells (PBMCs) in patients with proliferative capacity and antigen peptide-induced antigen-specific T Cell killing toxicity, and gradually identify HLA-A 觹 0201 restricted CD8 + CTL cell epitope. RESULTS: Five HLA-A 觹 0201-restricted antigen peptides were preliminarily predicted in five databases, of which two antigenic peptides (159-167aa and 165-174aa) showed higher levels of HLA-A 觹 0201 on T2 cells The two antigen peptides can stimulate PBMC proliferation in most HLA-A 觹 0201-positive PBC patients, and the CTLs induced by it are specifically cytotoxic. CONCLUSIONS: KLSEGDLLA POINT 159 ~ 167aa located on the PDC-E2 lactone domain) and LLAEIETDKA POINT 165 ~ 174aa are HLA-A 觹 0201-restricted CD8 + CTL epitopes in PBC patients.